Graves病遗传学研究进展

Graves病为多基因遗传病，是多对易感基因与环境因素共同作用的结果。本文就Graves病发病的遗传易感性、家族遗传方式、免疫遗传学特点以及今后的研究趋向等作一综述。     

Graves'病患者血清中IFN-γ测定的临床意义

近年来，围绕Graves’病的发病机理在分子生物学水平上进行了大量研究，取得一些进展，细胞因子在Graves’病发病中所起的作用受到越来越多的国内外专家的重视。我们通过动态观察Graves’病患者血清中γ-干扰素（interferon γ，IFN-γ）的含量的变化，探讨IFN-γ在Graves’病发病中的作用。     

Graves病遗传学研究进展

Graves病为多基因遗传病 ,是多对易感基因与环境因素共同作用的结果。本文就 Graves病发病的遗传易感性、家族遗传方式、免疫遗传学特点以及今后的研究趋向等作一综述     

TNFβ基因微卫星多态性与Graves病的相关性研究

目的 研究Graves病患者TNF(tumor necrosis factor)β基因第一内含子的微卫星多态性(TNFc)，分析TNFc与Graves病发病的关联，进一步探讨Graves病的发病机制。方法 选取Graves病实验组和正常对照组患者各36例，应用聚合酶链反应(PCR)技术，扩增具有微卫星多态性的TNFβ基因第一内合于，通过聚丙烯酰胺凝胶电泳，分析两组的基因频率和基因型频率。结果 TNFc微卫星多态性合有两个等位基因(TNFcl和TNFc2)及三种基因型(纯合于TNFclcl和TNFc2c2，亲合于TNFclc2)；Graves病实验组的TNFc2基因频率高于正常对照组，有显著性差异(x^2=4，02，P＜0．05)，TNFclcl基因型频率在Graves病实验组与正常对照组无显著性差异(X^2=2．72，P＞0．05)。结论 TNFc2等位基因与Graves病发病的易感性有关联，TNFclcl基因型在Graves病发病的遗传基础中不起重要作用。     

Graves病相关的易感基因研究进展

Graves病是一种器官特异性自身免疫病，是一种由多个基因与环境因素参与的多基因病。尽管进行了大量的研究, GD的遗传学机制至今尚未阐明。与GD发病相关的候选基因主要分成两大类：（1）与机体免疫调节有关的基因：（2）与甲状腺生理机能调节有关的基因。本文就与 GD易感性研究最多的基因 HLA 、CTLA-4 、TSH-R 及一些细胞因子作一简要综述。     

华东地区汉族人群IL12B基因多态性与Graves病相关性研究

目的 研究华东地区汉族人群IL12B基因3’UTR多态性与Graves病的相关性。方法 用直接测序法对华东地区93例Graves病患者及94例正常对照者的IL12B基因的3’UTR 的多态位点进行检测，分析华东地区汉族人群IL12B基因与Graves病的相关性。结果 在IL12B的3’UTR区检测到2个SNPs：1188A/C和1358-/G，基因型分布在GD患者和正常对照中无统计学差异；单倍型分析显示A-频率在GD患者中显著低于正常对照：P=0.046。结论 IL12B可能与GD的易感性有关。     

T细胞受体Vβ基因在自身免疫性甲状腺病病变组织中的表达及意义

目的研究国人Graves病、桥本甲状腺炎甲状腺内浸润T淋巴细胞的T细胞受体(TCR)Vβ基因家族的利用,发现优势利用基因,为阐明自身免疫性甲状腺病(AITD)的T细胞克隆的分布模式及防治提供依据.方法穿刺或手术取得12例Graves病、15例桥本甲状腺炎病人的甲状腺组织,提取RNA.抽取5例Graves病、5例桥本甲状腺炎和7例正常人之外周静脉血,分离外周血淋巴细胞(PBL),提取RNA.以24个Vβ基因家族特异的寡核苷酸为上游引物,以一个Cβ特异的寡核苷酸为下游引物,进行逆转录-聚合酶链反应,比较各个Vβ基因家族的表达.结果Graves病及桥本甲状腺炎患者甲状腺内TCRVβ基因家族的平均表达阳性数分别为(5.3±1.2)个,(13.4±3.0)个;而且Vβ3、Vβ5和Vβ8的表达在Graves病更常见.以上两组病人及正常对照PBL之TCRVβ基因家族的平均表达分别为(23.0±1.0)个,(22.2±1.3)个和(22.4±1.7)个.结论Graves病甲状腺内淋巴细胞的TCRVβ基因显示明显的限制性利用,某些Vβ家族的使用率更高,提示寡克隆扩增的甲状腺抗原特异性T细胞可能与Graves病的发病有关,这可能有重要的治疗意义.相反,桥本甲状腺炎病变内TCRVβ利用的限制性较差或无限制性,可能与随疾病进展,非特异性免疫机制的介入有关.     

Ⅱ型糖尿病的遗传学研究进展

遗传因素在Ⅱ型糖尿病(T2DM)的发生发展中起着重要作用。T2DM是一种多基因病，其遗传模式有主效基因、微效基因和单基因等模式。寻找T2DM易感基因的策略主要有基因组扫描、连锁分析和遗传关联研究。近年来，应用这些遗传学分析方法人们发现了一些与T2DM易感性关联的染色体区域和基因，并取得了长足的进展，在此简要加以综述。     

单核苷酸多态性与胃癌的关系研究进展

单核甘酸多态性(SNP)被称为第三代遗传标记,具有很多研究方法。单核甘酸多态性作为遗传标记在人类肿瘤、糖尿病等多基因病的研究中取得了一定的进展。本文综述了与胃癌相关的酶基因、细胞增殖相关基因、癌基因和抑癌基因、粘附分子基因和细胞因子基因的单核苷酸多态性,以分析不同基因型与胃癌的相关性。     

4p14区段和CTLA-4基因多态性与Graves病相关

目的探讨中国安徽蚌埠地区汉族人群4p14区段位点rs6832151和CTLA-4基因4个SNPs(单核苷酸多态性)位点基因多态性与Graves病相关性,和基因-基因交互作用对Graves病的影响。方法提取611例诊断明确的GD患者和644名健康对照者的全基因组DNA,用Taq Man探针技术进行基因分型,使用Plink和Haploview等统计软件分析这些位点与蚌埠地区汉族人群Graves病的相关性。结果 4p14区段位点rs6832151的等位基因、基因型频率在GD组和对照组之间有差异(P0.05),CTLA-4基因区域内rs231804和rs231726两个SNP位点基因型在显性模型下差异显著(P0.05);GMDR模型显示CTLA-4基因内rs10197319、rs231726、rs231804、rs1024161位点和4p14区段内rs6832151位点组成的五阶模型(P=0.001)为最佳模型,CTLA-4基因内rs1024161和rs10197319位点之间上位效应分析结果有差异(P0.05)结论 4p14区段rs6832151,CTLA-4基因区域内rs231804和rs231726位点基因多态性与蚌埠地区汉族人群Graves病的遗传易感性相关;rs6832151(4p14区段)和rs10197319、rs231726、rs231804、rs1024161(CTLA-4基因)5个SNP位点间的基因-基因交互作用与Graves病相关,CTLA-4基因内rs1024161和rs10197319位点之间存在上位效应。     

Studies on the structure and function of the apolipoprotein(a) gene

Lp(a) is an LDL-like lipoprotein that is a major inherited risk factor for atherosclerosis. It is distinguished from Lp(a) by the addition of apolipoprotein(a). The gene structure of apolipoprotein(a) is homologous to plasminogen, and competition with plasminogen activity may account for some of the pathophysiology associated with Lp(a). Six highly related genes have now been identified, and at least four are found in close proximity in overlapping genomic clones. Studies have begun on the regulation of apolipoprotein (a) gene expression, and the human apolipoprotein(a) gene has been inserted into transgenic mice, where it leads to the development of arterial lesions.     

Distribution of six neuropeptides in the nucleus tractus solitarii of the rat: an immunohistochemical analysis

Distribution of substance P-, [Leu]enkephalin-, cholecystokinin-8-, neurotensin-, avian pancreatic polypeptide- and gamma-melanocyte stimulating hormone-like immunoreactive structures were investigated in the nucleus tractus solitarii of the rat by means of the indirect immunofluorescence method. The density of the immunoreactive structures varied markedly according to neuropeptides or subnuclei, with the medial and commissural nuclei containing the highest density. This suggests that the peptides examined play a role in cardiovascular function. However, as seen in the substance P- and [Leu]enkephalin-like immunoreactive structures, these peptides were widely distributed in the nucleus tractus solitarii in addition to the commissural and medial nuclei; a high density of immunoreactive fibers in the ventral, dorsolateral and intermediate subnuclei. In addition to the immunoreactive fiber plexus, a group of immunoreactive cells was also identified in the subnuclei mentioned above. These findings strongly suggest that substance P- and [Leu]enkephalin-like immunoreactive structures are involved not only in cardiovascular function but also in other functions such as respiration, at least in the rat. Finally, the present study demonstrated that the area postrema, particularly its lateral portion, contains various neuropeptide-like structures, both neurons and fibers, substance P-, [Leu]enkephalin-, cholecystokinin-8- and neurotensin-like immunoreactive neurons and fibers, and avian pancreatic polypeptide- and gamma-melanocyte stimulating hormone-like immunoreactive fibers.     

Nystagmus induced by stimulation of the nucleus of the optic tract in the monkey

Summary 1. The nucleus of the optic tract (NOT) was electrically stimulated in alert rhesus monkeys. In darkness stimulation evoked horizontal nystagmus with ipsilateral slow phases, followed by after-nystagmus in the same direction. The rising time course of the slow phase velocity was similar to the slow rise in optokinetic nystagmus (OKN) and to the charge time of optokinetic after-nystagmus (OKAN). The maximum velocity of the steady state nystagmus was approximately the same as that of OKAN, and the falling time course of the after-nystagmus paralled OKAN. 2. Increases in frequency and duration of stimulation caused the rising and falling time constants of the nystagmus and after-nystagmus to become shorter. Changes in the falling time constant of the after-nystagmus were similar to changes in the time constant of OKAN produced by increases in the velocity or duration of optokinetic stimulation. 3. Stimulus-induced nystagmus was combined with OKN, OKAN and per- and post-rotatory nystagmus. The slow component of OKN as well as OKAN could be prolonged or blocked by stimulation, leaving the rapid component of OKN unaffected. Activity induced by electrical stimulation could also sum with activity arising in the semicircular canals to reduce or abolish post-rotatory nystagmus. 4. Positive stimulus sites for inducing nystagmus were located in the posterolateral pretectum. This included portions of NOT that lie in and around the brachium of the superior colliculus and adjacent regions of the dorsal terminal nucleus (DTN). 5. The data indicate that NOT stimulation had elicited the component of OKN which is responsible for the slow rise in slow phase velocity and for OKAN. The functional implication is that NOT, and possibly DTN, are major sources of visual information related to retinal slip in the animal's yaw plane for semicircular canal-related neurons in the vestibular nuclei. Analyzed in terms of a model of OKN and OKAN (Cohen et al. 1977; Waespe et al. 1983), the indirect pathway, which excites the velocity storage mechanism in the vestibular system to produce the slow component of OKN and OKAN, lies in NOT in the monkey, as it probably also does in cat, rat and rabbit. Pathways carrying activity for the rapid rise in slow phase velocity during OKN or for ocular pursuit appear to lie outside NOT.Supported by NIH grants EY02296, EY04148, EY01867 and PSC-CUNY FRAP award 6-63231     

脑功能多种成像方式整合技术的研究及其进展

近年来,人们运用脑的局部血流技术对人体大脑功能的研究发展迅速,尤其是PET和f MRI具有极高的空间分辨率,但时间分辨率跟不上大脑神经生理变化的速度,与之相对应的EEG和MEG能够以毫秒级的速度跟踪神经元活动产生的电信号,但空间分辨率低,因此,为了获取大脑神经元活动的高时一空动态图像,需要对多种方法获得的信号进行整合。本文阐述了脑功能成像技术的物理和生理学基础,讨论了脑功能多种成像方式整合技术的各种方法,并对该领域的发展方向和所面临的挑战进行了简要的分析。     

仿生型BG-COL-HYA-PS复合支架细胞相容性

目的:研究仿生型BG-COL-PS-HYA复合支架材料与成骨细胞的相容性。方法:将小鼠胚胎成骨细胞种植于BG-COL-HYA-PS、BG-COL复合材料、58SBG支架上,用MTT法、ALP活性测定等观察细胞在材料中的生长情况。通过体外实验方法,观察其生物相容性。结果:成骨细胞在BG-COL-HYA-PS材料上能良好粘附、增殖,而在58SBG材料上粘附差、细胞逐渐死亡。MTT法结果显示:联合培养后,BG-COL-HYA-PS组的OD值为0.314±0.004,5天时达到0.621±0.002,分别为58SBG组的1.49倍和1.44倍,P<0.05。结论:仿生型BG-COL-PS-HYA复合支架具有天然骨分级结构和有良好的生物相容性,在诱导成骨细胞增殖方面性能优越,可作为骨组织工程支架材料,临床应用前景广阔。     

粉防己碱对戊四唑致(癎)大鼠皮层电图和海马超微结构的影响

目的:探讨钙拮抗剂粉防己碱(tetradrine,Tet)对戊四唑(Pentylentetrazol,PTZ)诱导的大鼠癫(癎)模型的作用.方法:健康SD大鼠24只,随机分为4组,对照组和Tet 3个剂量组(15 mg/kg,30 mg/kg,60 mg/kg)各6只.观察大鼠腹腔注射PTZ前后癫(癎)发作的情况,按Racine分级标准分级,同时记录皮层电图(ECoG),观察PTZ注射后到出现(癎)样放电的潜伏期及1 h内(癎)样放电累加的持续时间.电镜观察海马神经元超微结构的改变.结果:对照组给PTZ后均出现癫(癎)发作,程度均为5级,Tet组发作程度明显减轻.ECoG上出现(癎)样放电的潜伏期延长,(癎)样放电在1 h中的持续时间缩短.同时,Tet也能明显减轻PTZ致(癎)大鼠海马CA1区锥体细胞线粒体的损伤程度.结论:Tet对PTZ诱导的癫(癎)大鼠的发作有明显的对抗作用,对海马锥体神经元的拟伤具有保护作用.     

YAP is up-regulated in the bronchial airway smooth muscle of the chronic asthma mouse model

Asthma is characterized by leukocytic infiltration and tissue remodeling with structural changes including subepithelial fibrosis and ASM cells proliferation. The Hippo pathway is a key regulatory point involved in cell proliferation, fibroblasts, and smooth muscle cell differentiation. In order to disclose the relation between asthma and the Hippo pathway, expression of the Yes-associated protein (YAP), a key gene in the Hippo pathway, in the bronchial smooth muscle of chronic asthma model (CAM) was studied. 40 mice were randomly divided into control (wide type) and experimental group to construct CAM using chicken ovalbumin (OVA). Pathological changes of the lung tissues were observed in the CAM mice compared with the control using HE staining method. Immunohistochemistry (IHC) was used to detect if YAP protein is expressed in the lung tissues. The pathological changes of the CAM group showed that a large number of inflammatory cells infiltration including mainly lymphocytes and a small amount of eosinophilic, with the presence of certain airway smooth muscle hyperplasia, was observed in comparison with the control. IHC results showed that the YAP protein was significantly increased compared with the control groups (P < 0.01). This result was further confirmed by quantitative real-time PCR (qPCR) assay which detected the up-regulation of the YAP gene (P < 0.01) and Western blot. In conclusion, the YAP protein was significantly expressed in the bronchial airway tissues of the CAM mice, and could be used as an indicator for asthma.     

缺氧缺血性脑病患儿NO、NOS水平与脂质过氧化关系的研究

目的:探讨了缺氧缺血性脑病患儿血清NO、NOS水平与脂质过氧化的关系.方法:应用生化法对30例缺氧缺血性脑病患儿进行了血清NO、NOS测定和化学比色法测定血清丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-PX)含量,并以35名正常健康人作比较.结果:缺氧缺血性脑病患儿血清NO、SOD、GSH-PX水平显著低于正常人组(P＜0.01),而NOS、MDA水平则显著高于正常人组(P＜0.01),相关分析显示:血清NO、SOD、GSH-PX水平与MDA水平呈明显负相关(r=-0.4286、-0.4125、-0.4108,P＜0.05).结论:缺氧缺血性脑病患儿血清NO、SOD、GSH-PX水平的降低与活性代谢紊乱密切相关.     

Peptide domains involved in the localization of the porcine reproductive and respiratory syndrome virus nucleocapsid protein to the nucleolus

The nucleocapsid (N) protein of porcine reproductive and respiratory syndrome virus (PRRSV) is the principal component of the viral nucleocapsid and localizes to the nucleolus. Peptide sequence analysis of the N protein of several North American isolates identified two potential nuclear localization signal (NLS) sequences located at amino acids 10-13 and 41-42, which were labeled NLS-1 and NLS-2, respectively. Peptides containing NLS-1 or NLS-2 were sufficient to accumulate enhanced green fluorescent protein (EGFP) in the nucleus. The inactivation of NLS-1 by site-directed mutagenesis or the deletion of the first 14 amino acids did not affect N protein localization to the nucleolus. The substitution of key lysine residues with uncharged amino acids in NLS-2 blocked nuclear/nucleolar localization. Site-directed mutagenesis within NLS-2 identified the sequence, KKNKK, as forming the core localization domain within NLS-2. Using an in vitro pull-down assay, the N protein was able to bind importin-alpha, importin-beta nuclear transport proteins. The localization pattern of N-EGFP fusion peptides represented by a series of deletions from the C- and N-terminal ends of the N protein identified a region covering amino acids 41-72, which contained a nucleolar localization signal (NoLS) sequence. The 41-72 N peptide when fused to EGFP mimicked the nucleolar-cytoplasmic distribution of native N. These results identify a single NLS involved in the transport of N from the cytoplasm and into nucleus. An additional peptide sequence, overlapping NLS-2, is involved in the further targeting of N to the nucleolus.     

Effects of lesions of the nucleus of the optic tract on optokinetic nystagmus and after-nystagmus in the monkey

Summary 1. The nucleus of the optic tract (NOT) and the dorsal terminal nucleus (DTN) of the accessory optic system were lesioned electrolytically or with kainic acid in rhesus monkeys. When lesions involved NOT and DTN, peak velocities of optokinetic nystagmus (OKN) with slow phases toward the side of the lesion were reduced, and optokinetic after-nystagmus (OKAN) was reduced or abolished. The jump in slow phase eye velocity at the onset of OKN was smaller in most animals, but was not lost. Initially, there was spontaneous nystagmus with contralateral slow phases. OKN and OKAN with contralateral slow phases were unaffected. 2. Damage to adjacent regions had no effect on OKN or OKAN with two exceptions: 1. A vascular lesion in the MRF, medial to NOT and adjacent to the central gray matter, caused a transient loss of the initial jump in OKN. The slow rise in slow phase velocity was prolonged, but the gain of OKAN was unaffected. There was no effect after a kainic acid lesion in this region in another animal. 2. Lesions of the fiber tract in the pulvinar that inputs to the brachium of the superior colliculus caused a transient reduction in the buildup and peak velocity of OKN and OKAN. 3. In terms of a previous model (Cohen et al. 1977; Waespe et al. 1983), the findings suggest that the indirect pathway that activates the velocity storage integrator in the vestibular system to produce the slow rise in ipsilateral OKN and OKAN, lies in NOT and DTN. Activity for the rapid rise in OKN, carried in the direct pathway, is probably transmitted to the pontine nuclei and flocculus via an anatomically separate fiber path-way that lies in the MRF. A fiber tract in the pulvinar that inputs to the brachium of the superior colliculus appears to carry activity related to retinal slip from the visual cortex to NOT and DTN.Abbreviations used in Figures BIC brachium of the inferior colliculus - BSC brachium of the superior colliculus - C caudate nucleus - CG central gray - CL Centralis lateralis - dbc decussation of the brachium conjunctivum - DTN dorsal terminal nucleus of the accessory optic system - IC inferior colliculus - Hb habenular nucleus - hc habenular commissure - LD lateralis dorsalis - LGn lateral geniculate nucleus - MD medialis dorsalis - MGn medial geniculate nucleus - MLF median longitudinal fasciculus - MRF mesencephalic reticular formation - cMRF central mesencephalic reticular formation - NL nucleus limitans - NLL nucleus of the lateral lemniscus - NOT nucleus of the optic tract - PB parabigeminal nucleus - pc posterior commissure - Pi pineal gland - PON pretectal olivary nucleus - Pt pretectum - Pulv pulvinar - R nucleus reticularis - RN red nucleus - RpN raphe nucleus - RTP nucleus reticularis tegmenti pontis - SC superior colliculus - SCpit superior cerebellar peduncle - VPL ventralis postero-lateralis - VPM ventralis posteromedialis - III oculomotor nucleus - IV trochlear nucleus - IVn trochlear nerve - Vm mesencephalic trigeminal nucleus